武夷黑猪与杜洛克猪HSPB1基因克隆、生物信息学对比分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2025.09.023

摘要/Abstract

摘要： 【目的】对比分析武夷黑猪与杜洛克猪热休克蛋白B1(heat shock protein beta-1,HSPB1)结构和功能，为研究动物机体抗应激机制提供参考。【方法】通过克隆武夷黑猪和杜洛克猪HSPB1基因并测序，同时运用生物信息学方法比对相似性，构建系统进化树，对比分析两者HSPB1蛋白的特性、结构和功能的差异，以及在两个猪种中各组织的表达水平。【结果】成功克隆武夷黑猪与杜洛克猪HSPB1基因，编码区均为624 bp，均共编码207个氨基酸，两者HSPB1核苷酸和氨基酸相似性分别为99.0%和98.6%，存在123^G→T、233^T→C核苷酸位点的变异，同时引起41^E→D、78^L→P氨基酸的差异。核苷酸序列进化树显示，两者均与蝙蝠亲缘关系最近；氨基酸序列进化树显示，两者均与羊亲缘关系最近。两者有相同的基因启动子与转录因子，在蛋白质的信号肽、跨膜区、保守基序、理化性质、亲/疏水性等特性上保持一致。进一步对比分析发现，该蛋白中存在1个仅可被糖原合酶激酶-3(glycogen synthase kinase-3，GSK-3)所识别的新的磷酸化位点，和4个可被化学因子结合作用的差异位点。武夷黑猪HSPB1蛋白在线粒体中占比比杜洛克猪少，氨基酸序列的二级结构、三级结构存在7个区域的差别。虽然两者都与磷酸化激酶互作密切，但武夷黑猪比杜洛克猪HSPB1蛋白多1个细胞骨架功能条目。HSPB1基因在两个猪种各组织中广泛表达，且品种间均无显著差异，其中都在背最长肌中表达量最高。【结论】成功扩增武夷黑猪与杜洛克猪HSPB1基因。相较于杜洛克猪，武夷黑猪HSPB1在细胞质中含量更高，且存在磷酸化位点和化学因子结合位点的差异。但两者在相同组织中的表达量无差异，都在背最长肌中表达量最高。结果为深入研究猪源HSPB1在抗应激反应中发挥的作用提供了理论基础。

关键词: 武夷黑猪; 杜洛克猪; HSPB1基因; 生物信息学; 组织表达

Abstract: 【Objective】 The purpose of this study was to compare and analyze the similarities and differences in the structure and function of heat shock protein beta-1 (HSPB1)between Wuyi Black pig and Duroc pig,and provide a reference for studying animal anti-stress mechanism. 【Method】 The HSPB1 gene of Wuyi Black pig and Duroc pig was cloned and sequenced,and bioinformatics methods were used to compare similarities.A phylogenetic tree was constructed.And a comparative analysis was conducted on the differences in the characteristics,structure,and function of HSPB1 protein between the two pig breeds,as well as the expression of each tissue in the two breeds. 【Result】 The HSPB1 gene was successfully cloned from both Wuyi Black pig and Duroc pig.The coding sequence (CDS) of each gene was 624 bp in length,encoding a protein composed of 207 amino acids.The nucleotide and amino acid sequence similarities between the two breeds were 99.0% and 98.6%,respectively，with variations in nucleotide sites 123^G→T and 233^T→C,as well as differences in amino acids 41^E→D and 78^L→P.The nucleotide sequence evolutionary tree showed that both were closely related to Desmodus rotundus;The evolutionary tree of amino acid sequences showed that both were closely related to Ovis aries.Both had the same gene promoters and transcription factor binding sites,and their HSPB1 proteins displayed consistent features in terms of signal peptides,transmembrane regions,conserved motifs,physicochemical properties,and hydrophilicity/hydrophobicity profiles.Further comparative analysis identified one novel phosphorylation site uniquely recognized by glycogen synthase kinase-3 (GSK-3) and four differential sites potentially involved in interactions with chemical effectors.However,the mitochondrial proportion of HSPB1 protein in Wuyi Black pig was lower than that in Duroc pig.Moreover,seven distinct regions of secondary and tertiary structural divergence were observed between the two proteins.Although both HSPB1 proteins exhibited strong interactions with phosphorylation-related kinases,the Wuyi Black pig had an additional functional annotation related to cytoskeletal organization. HSPB1 gene was broadly expressed across various tissues in both pig breeds,with no significant differences between breeds.In both breeds,the highest expression was observed in the longissimus dorsi muscle. 【Conclusion】 HSPB1 genes of Wuyi Black pig and Duroc pig were successfully amplified.Compared with Duroc pig, HSPB1 in Wuyi Black pig showed higher cytoplasmic content and differences in phosphorylation and chemical factor binding sites.However,the expression was found to be similar across corresponding tissues in both breeds,with the highest expression in longissimus dorsi muscle.The results provided a theoretical foundation for further studying the specific role of porcine HSPB1 in anti-stress response.

Key words: Wuyi Black pig; Duroc pig; HSPB1 gene; bioinformatics; tissue expression

中图分类号:

S828
Q78

李指全, 高萌若, 温逸俊, 傅思静, 徐舒平, 杨涛涛, 张志榜, 方霞, 李凯, 李鹏成. 武夷黑猪与杜洛克猪HSPB1基因克隆、生物信息学对比分析及组织表达研究[J]. 中国畜牧兽医, 2025, 52(9): 4248-4259.

LI Zhiquan, GAO Mengruo, WEN Yijun, FU Sijing, XU Shuping, YANG Taotao, ZHANG Zhibang, FANG Xia, LI Kai, LI Pengcheng. Cloning,Bioinformatics Comparative Analysis and Tissue Expression of HSPB1 Gene Between Wuyi Black Pig and Duroc Pig[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4248-4259.

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